The present invention relates to the detection and treatment of breast disease.
Breast cancer is one of the largest classes of malignant disease in women. However, breast cancer presents inherent difficulties in regard to the ease with which it is detected and diagnosed. This is in contrast to detection of some other common cancers, including skin and cervical cancers, the latter of which is based on cytomorphologic screening techniques.
Early detection of breast cancer represents a compelling goal in oncology. Although techniques such as computerized tomography, mammography, and magnetic resonance imaging have greatly improved tumor surveillance over the past decade, there still remains a need for serologic and other blood-based assays.
Serologic assays are easily performed, inexpensive, and analytically-sensitive and can be serially run over time with relative ease. The essence of breast cancer screening, using tumor marker detection, is to efficiently identify a group of higher-risk individuals from within a large population. Thereafter, confirmatory testing is implemented to establish a diagnosis of malignancy.
There are several classifications of tumor markers possible, based upon the structure or biological function of the marker. Tumor marker classifications include tissue specific antigens (e.g., PSA, NSE, PAP, calcitonin, HCG), major histocompatibility complex (xe2x80x9cMHCxe2x80x9d) antigens, viral antigens (e.g., HTLV-I gag protein), oncogene products (e.g., c-HER-2/Neu), oncofetal markers (e.g., CEA, AFP), hormones (e.g., thyroid hormones), enzymes (e.g., telomerase, galactosyltransferase), and altered glycoproteins/glycolipids (e.g., polymorphic epithelial mucins). It should be noted that these classification schemes are imprecise and contain redundancies. For example, calcitonin is an important serological marker for medullary carcinoma of the thyroid and may be classified not only as a hormone but also as a tissue specific protein of the thyroid. Likewise, PSA, HCG, thyroid hormones, PAP, and NSE are tissue specific proteins and also exhibit enzymatic or hormonal activities. Generally, tumor markers providing high clinical utility reside in the broadly defined tissue specific class. This class of tumor markers contains enzymes, isoenzymes, hormones, growth factors, and other molecules with biologic activity.
The importance of a tumor marker""s being tissue specific is illustrated by one of the best known tumor antigens, carcinoembryonic antigen (xe2x80x9cCEAxe2x80x9d). When first discovered, CEA was thought to be specific to cancers of the digestive system. However, CEA has since been detected in normal adults as well as in patients with benign liver disease, such as alcoholic hepatitis or biliary obstruction. Because of the overall lack of specificity and sensitivity, there being no threshold difference in CEA levels that serves to separate benign from malignant conditions, CEA cannot be used in a general diagnostic test. Instead, it is principally used to monitor a patient""s response to treatment.
To be useful in serologic assays, a tumor marker should be one that is released into the bloodstream as a circulating marker. Circulating antigens are now known to exist in breast cancer. Breast tissue markers, such as casein (Franchimont et al., Cancer, 39:2806-2812 (1977)) and xcex1-lactalbumin (Kleinberg et al., Science, 190:276-278 (1975)) and purported cancer markers, such as glycosyl transferases (Ip et al., Cancer Res., 38:723-728 (1978) and Dao et al., J. Natl. Cancer Inst., 65:529-534 (1980)), glycolipids (Kloppel et al., Proc. Natl. Acad. Sci. USA, 74:3011-3013 (1977)), and phospholipids (Skipski et al., Proc. Soc. Exp. Biol. Med., 136:1261-1264 (1971)) have all been used in various diagnostic techniques for breast cancer but have not gained widespread acceptance as breast cancer markers. More recently, circulating human mammary epithelial antigens have been proposed as specific markers for breast cancer (Ceriani et al., Proc. Natl. Acad. Sci. USA, 79:5420-5424 (1982)). Burchell et al., Int. J. Cancer, 34:763-768 (1984) describes monoclonal antibodies which detect high molecular weight mucin-like antigens elevated in patient serum. Hayes, J. Clin. Invest., 75:1671-1678 (1985) also describes a monoclonal antibody that recognizes a high molecular weight mammary epithelial antigen present in elevated amounts in the plasma of breast cancer patients. See also Papsidero et al., Cancer Res., 44:4653-4657 (1984) and Taylor-Papadimitriou et al., Int. J. Cancer., 28:17-28 (1981). Other breast tissue specific proteins or markers include alpha, beta, and kappa caseins, alpha-lactalbumin, lactoferrin, and selected epithelial membrane antigens. These are described in Cohen et al., Cancer, 60:1294-1298 (1987); Bartkova, Eur. J. Cancer Clin. Oncol., 23:1557-1563 (1987); Weir et al., Cancer Detect. Prev., 4:193-204 (1981); de Almeida et al., Breast Cancer Res. Treat., 21:201-210 (1992); Skilton et al., Tumor Biol., 11:20-38 (1990); Earl et al., Cancer Res., 49:6070-6076 (1989); Barry et al., Amer. J. Clin. Path., 82:582-585 (1984); and Watson et al., Cancer Res., 56:860-865 (1996). None of these previously described antigens has been used as a basis for a widely accepted breast cancer clinical assay.
There have also been several attempts to develop improved methods of breast cancer detection and diagnosis based on oncogene mutations, gene amplification, and loss of heterozygosity in invasive breast cancer. These methods have not gained wide acceptance.
Despite the use of mammography and the development of some breast tissue specific markers, there still remains a need for simple and rapid methods for detecting breast cancer. The present invention is directed to meeting this need.
One aspect of the present invention relates to an isolated chemokine that is preferentially expressed in breast tissue or which can be detected in breast milk. The isolated chemokine includes about from about 100 to about 132 amino acids, has a deduced molecular weight of from about 10 to about 16 kDa, and has a deduced isoionic point of from about pH 10.1 to about pH 10.7.
The present invention also relates to peptides having an amino acid sequence corresponding to an antigenic portion of the subject chemokine, to antibodies which recognize this chemokine, and to isolated nucleic acid molecules which encode this chemokine.
The present invention also relates to an isolated nucleic acid molecule which, under stringent conditions, hybridizes to a nucleic acid molecule encoding a chemokine of the present invention or to a complement thereof.
In another aspect thereof, the present invention relates to an isolated nucleic acid molecule which encodes for a chemokine of the present invention.
The present invention also relates to a method for detecting breast disease in a patient. A sample of tissue or body fluid from the patient is contacted with a nucleic acid primer which, under stringent conditions, hybridizes to a nucleic acid molecule encoding a chemokine of the present invention or to a complement thereof. The sample of tissue or body fluid from the patient in contact with the nucleic acid primer is treated under conditions effective to amplify breast tissue specific nucleic acid molecules. The method further includes detecting the breast tissue specific nucleic acid molecules.
The present invention also relates to another method of detecting breast disease in a patient. In this method, a sample of tissue or body fluid from the patient is contacted with a nucleic acid probe under conditions effective to permit formation of a hybridization complex between the probe and breast tissue specific nucleic acid molecules. The nucleic acid probe is one which, under stringent conditions, hybridizes to a nucleic acid molecule encoding a chemokine of the present invention or to a complement thereof. The method further includes detecting the hybridization complex.
The present invention also relates to yet another method of detecting breast disease in a patient. The method includes providing an antibody or binding portion thereof which recognizes a chemokine of the present invention. The antibody or binding portion thereof is contacted with a liquid or tissue sample from the patient under conditions effective to permit binding of the antibody or binding portion thereof to the chemokine in the liquid or tissue sample. The method further includes detecting presence of antibody or binding portion thereof bound to the chemokine in the liquid or tissue sample.
The present invention, in another aspect thereof, relates to a method of treating breast disease in a patient. The method includes administering to the patient an effective amount of an antibody or binding portion thereof which recognizes a chemokine of the present invention.
The present invention also relates to another method of treating breast disease in a patient. The method includes administering to the patient an effective amount of a peptide which binds to a cellular receptor for a chemokine of the present invention.
The present invention also relates to a method of vaccinating a patient against breast disease. The method includes administering to the patient an effective amount of an antigenic portion of a chemokine of the present invention.
The chemokines, peptides, antibodies, and nucleic acid molecules of the present invention are useful in the early detection of various pathological states of the mammary gland, such as inflammations, infections, benign hyperplasias, and malignancies. In particular, they can be used in the early detection of breast cancer as well as for monitoring the presence or absence of metastatic breast cancer cells in a patient""s tissues and fluids, such as blood, lymph nodes, bone marrow, and other sites of disease dissemination. They can also be used to stage patients with breast cancer and to assess the effects of conventional breast cancer therapies. Furthermore, the chemokines, peptides, and antibodies of the present invention can be used to treat or prevent breast disease.